Production of a recombinant imported fire ant venom allergen, Sol i 2, in native and immunoreactive form  Margit Schmidt, PhDa, b, Thomas J. McConnell, PhDc, Donald R. Hoffman, PhDb  Journal of Allergy and Clinical Immunology  Volume 98, Issue 1, Pages 82-88 (July 1996) DOI: 10.1016/S0091-6749(96)70229-4 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Isolation of rSol i 2 from concentrated diafiltered supernatant of recombinant baculovirus grown in High-Five cells in Ex-Cell 400 medium on a Super-ODS reverse-phase column at 50° C with a gradient of acetonitrile in water containing TFA as an ion-pairing agent. The fraction from 11.00 to 11.50 minutes was collected, corresponding to the elution position of nSol i 2. Journal of Allergy and Clinical Immunology 1996 98, 82-88DOI: (10.1016/S0091-6749(96)70229-4) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 Silver-stained SDS-PAGE gel of culture supernatants from Sol i 2 recombinant baculovirus grown in Sf9 cells. Lanes from left to right are: n, natural Sol i 2; G, rSol i 2 baculovirus grown in Grace’s medium; Gm, Grace’s medium without serum; Mc, Sf9 cells grown in Sf-900 II SFM; 2, 10, and 13, clones 2, 10, and 13 of rSol i 2 baculovirus in Sf9 cells grown in Sf-900 II SFM. Journal of Allergy and Clinical Immunology 1996 98, 82-88DOI: (10.1016/S0091-6749(96)70229-4) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Dot immunoblots of natural Sol i 2 and rSol i 2 with monoclonal antibodies G260 and A4. Lanes 1, 2, and 3 are natural Sol i 2; lanes 4, 5, and 6 are rSol i 2. Dots 1 and 4 are untreated, dots 2 and 5 are boiled in SDS, and dots 3 and 6 are boiled in SDS in the presence of dithiothreitol. Journal of Allergy and Clinical Immunology 1996 98, 82-88DOI: (10.1016/S0091-6749(96)70229-4) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 A, Silver-stained SDS-PAGE gel of culture supernatants from two flasks of recombinant baculovirus grown in High-Five cells. Lanes are labeled n for natural Sol i 2; 1,2 for supernatants from rSol i 2 baculovirus; and c for control cells grown in Ex-Cell 400 medium. B, Silver-stained SDS-PAGE gel of purified natural (n) and recombinant (r) Sol i 2, each at three different concentrations. Journal of Allergy and Clinical Immunology 1996 98, 82-88DOI: (10.1016/S0091-6749(96)70229-4) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 A, Silver-stained SDS-PAGE gel of culture supernatants from two flasks of recombinant baculovirus grown in High-Five cells. Lanes are labeled n for natural Sol i 2; 1,2 for supernatants from rSol i 2 baculovirus; and c for control cells grown in Ex-Cell 400 medium. B, Silver-stained SDS-PAGE gel of purified natural (n) and recombinant (r) Sol i 2, each at three different concentrations. Journal of Allergy and Clinical Immunology 1996 98, 82-88DOI: (10.1016/S0091-6749(96)70229-4) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Plot of RAST specific binding of sera from 24 patients allergic to imported fire ant venom to natural Sol i 2 and rSol i 2. Linear regression coefficient is r = 0.94, with a slope of 1.08. Journal of Allergy and Clinical Immunology 1996 98, 82-88DOI: (10.1016/S0091-6749(96)70229-4) Copyright © 1996 Mosby, Inc. Terms and Conditions